CELL CYCLE REGULATION OF DNA REPAIR

DNA 修复的细胞周期调节

• 批准号: 6271474
• 负责人: Pablo Arenaz
• 金额: $ 9.07万
• 依托单位: UNIVERSITY OF TEXAS EL PASO
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-06-01 至 1999-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6271474
• 关键词: Bloom syndrome; CHO cells; DNA directed DNA polymerase; DNA repair; DNA replication; animal genetic material tag; cancer risk; cell cycle; endonuclease; enzyme activity; gene expression; gene mutation; genetic disorder; genetic library; genetic regulation; glycosidases; human genetic material tag; messenger RNA; molecular cloning; molecular pathology; northern blottings; nucleic acid sequence; polymerase chain reaction; transfection

The overall objective(s) of the proposed research is to gain a fundamental understanding of the mechanism(s) involved in the temporal regulation of the expression of DNA repair genes. A thorough analysis of the regulatory mechanisms involved in the regulation of DNA repair processes will increase our understanding of cell cycle mediated gene regulation in eukaryotes and our knowledge of the strategies cell use to ensure self perpetuation without error. The information gained on understanding the regulation of the expression of DNA repair processes should provide insight into the relationship between DNA repair and genetic diseases in which predisposition to cancer is a phenotypic characteristic. The temporal modulation of DNA repair processes appears to be a common feature in mammalian cells of different origins. In addition, there appears to be a correlation between altered cell cycle control of DNA repair processes and DNA repair deficiency or hypersensitivity to damage. However, the exact mechanism of this modulation is unclear. Several questions remain unanswered: Is the modulation at the level of transcription or post transcriptional? Is control initiated at translation or post translational modification; Alternatively, it could be a combination of all of the above. It is clear though, that both human and CHO cells will provide good models for understanding the regulatory processes involved. As part of a continuing effort to understand the regulatory mechanism(s) involved in the temporal modulation of DNA repair processes as well as the molecular basis for the genetic defect in Bloom's syndrome and repair deficient CHO cell lines, it is proposed to examine the control of DNA repair processes at the molecular level. It is planned to look at control mechanisms from several different aspects; 1) To examine whether or not control is at the level of transcription. It is planned to quantitate mRNA levels of various DNA repair genes throughout the cell cycle using Northern Blot hybridization; 2) To clone and sequence the major apurinic/apyrimidinic (AP) endonuclease from Chinese hamster cells. It is planned to determine if there are any DNA sequence irregularities between the normal and the repair deficient AP endonuclease which could explain the observed difference in temporal modulation; 3) To construct a genomic library from CHO cells. The resulting library will be screened for uracil DNA glycosylase (UDG) and AP endonuclease genes to look for common regulatory regions which may explain the coordinate control of these gene products; 4) Student training is an integral component of the proposed research. Graduate students will be given the opportunity to be trained in modern molecular biology techniques while developing an appreciation for the rigors of scientific endeavor.

拟议研究的总体目标是获得基本 了解与时间调节的机制 DNA修复基因的表达。 对监管的详尽分析 调节DNA修复过程的机制将增加 我们对真核生物中细胞周期介导的基因调节的理解和 我们对策略的了解，以确保自我永久存在 没有错误。 了解有关了解调节的信息 DNA修复过程的表达应提供对 DNA修复与遗传疾病之间的关系 癌症的易感性是表型特征。 DNA修复过程的时间调制似乎很常见 不同起源的哺乳动物细胞中的特征。 另外，那里 DNA的细胞周期控制改变似乎是一个相关性 修复过程和DNA修复缺乏或对损伤的过敏性。 但是，此调制的确切机制尚不清楚。 一些 问题仍未得到解决：是否在 转录还是转录后？ 是在翻译时启动的控制 或发布翻译修改； 或者，它可能是 上述所有结合。 但是很明显，人类和 CHO单元将提供理解调节的良好模型 涉及的过程。 作为不断努力的一部分，以了解监管机制 参与DNA修复过程的时间调制以及 Bloom综合征和修复的遗传缺陷的分子基础 缺乏CHO细胞系，建议检查DNA的控制 修复分子水平的过程。 计划查看控制 来自几个不同方面的机制； 1）检查是否 控制处于转录水平。 计划定量mRNA 整个细胞周期的各种DNA修复基因的水平使用北部 印迹杂交； 2）克隆并序列专业 来自中国仓鼠细胞的肾上腺素/肾上腺素（AP）核酸内切酶。 这是 计划确定之间是否存在任何DNA序列不规则性 正常和维修缺乏AP核酸内切酶，这可以解释 观察到时间调制的差异； 3）构建基因组 来自Cho细胞的库。 最终的库将筛选为uracil DNA糖基酶（UDG）和AP内切核酸酶基因寻找常见 可以解释这些基因的坐标控制的调节区域 产品; 4）学生培训是拟议的组成部分 研究。研究生将有机会接受培训 现代分子生物学技术，同时对 科学努力的严格。